Choline oxidase: a useful tool for high-throughput assays of acetylcholinesterase phospholipase D phosphatidylcholine-specific phospholipase C and sphingomyelinase

摘要

Abstract: Choline oxidase converts one molecule of choline to generate two molecules of H$-2$/O$-2$/. In a 1:1 reaction stoichiometry, horseradish peroxidase-catalyzed oxidation of the nonfluorescent and colorless H$-2$/O$-2$/ indicator, the Amplex Red reagent, by H$-2$/O$-2$/ produces the highly fluorescent resorufin, which has an excitation maximum at 563 nm and emission maximum at 587 nm. Using the Amplex Red reagent as an H$-2$/O$-2$/ indicator, assays of phospholipase D, phosphatidylcholine-specific phospholipase C, acetylcholine, acetylcholinesterase, and sphingomyelinase have been developed in choline oxidase- and HRP-coupled reactions. These assays have been developed in a microplate with a one-step procedure.!9